High-altitude Andean wetlands are pristine environments with extreme conditions such as high UV radiation, high heavy metal content (mainly arsenic), high salinity, and oligotrophy. In this paper, the UV-B resistance and tolerance to arsenic of phylogenetically characterized bacteria (Actinobacteria [six isolates], Firmicutes [four isolates], and gamma-Proteobacteria [three isolates]) isolated from Laguna Vilama (4400-m altitude) and Laguna Azul (4560 m) were determined. In addition, given that multiple antibiotic resistances were also determined, a relationship between antibiotic resistances as a consequence of mutagenic ability or in relation to metal resistance is proposed. High UV-B resistances were found, since after 30 min (0.7 KJ m(-2)) and 60 min (1.4 KJ m(-2)) of irradiation, most of the studied bacteria did not show a decreased survival; what is more, many of them had an improved survival with the increased doses. Augmentations in mutagenesis rates were observed after UV-B irradiation in only 4 of the 13 tested isolates. Arsenite tolerance was also established in 8 of the 13 tested strains: Staphylococcus saprophyticus A3 and Micrococcus sp. A7, which were able to grow in media containing up to 10 mM As(III). Finally, predominance of antibiotic resistances (azithromycin, erythromycin, clarithromycin, roxithromycin, streptomycin, chloramphenicol, gentamycin, kanamycin, tetracycline, and ampicillin) was found, in all the isolated strains from both wetlands, with unexpectedly high minimal inhibitory concentrations (MICs; >2 mg mL(-1)) for macrolides. These results demonstrate that in extreme environments like high-altitude wetlands there is a correlation of multiresistances to UV-B radiation and arsenic, and that antibiotic resistances are also widespread in these pristine environments, where antibiotic selective pressure is supposed to be absent.
Azospirillum brasilense (strains REC3, RLC1, PEC5) were root inoculated in strawberry plants of the cultivars 'Milsei', 'Selva' and 'Camarosa' to assess plant growth-promoting effects. The bacteria were able to promote plant growth (expressed as root length, root area, and dry weight of root and shoot), depending on the genotypes of plants and bacteria used, whereas the stolon production (3-4) depended only on the strawberry cultivar. To explain whether root exudates plays any role on the growth-promotion observed herein, total protein and sugar were determined, and chemotaxis properties were evaluated. The strains showed positive chemotaxis toward the root exudates, being influenced by the total sugars content, suggesting that the latter plays an important role in the chemotaxis effect and may contribute to enhance the root capacity to recruit azospirilla from rhizosphere, thus improving the growth-promoting effect exerted by these bacteria.
Azospirillum species are free-living nitrogen-fixing bacteria commonly found in soil and in association with roots of different plant species. For their capacity to stimulate growth they are known as plant growth-promoting bacteria (PGPB). In this work, we demonstrate the natural occurrence and colonization of different parts of strawberry plants by Azospirillum brasilense in the cropping area of Tucumán, Argentina. Although bacteria isolations were carried out from two strawberry cultivars, e.g., Camarosa and Pájaro, attempts were successful only with the cultivar Camarosa. Whereas different strains of Azospirillum were isolated from the root surface and inner tissues of roots and stolons of the cultivar Camarosa, we have not obtained Azospirillum isolates from the cultivar Pájaro. After microbiological and molecular characterization (ARDRA) we determined that the isolates belonged to the species A. brasilense. All isolates showed to have the capacity to fix nitrogen, to produce siderophores and indoles. Local isolates exhibited different yields of indoles production when growing in N-free NFb semisolid media supplemented or not with tryptophan (0.1 mg ml À1 ). This is the first report on the natural occurrence of A. brasilense in strawberry plants, especially colonizing inner tissues of stolons, as well as roots. The local isolates showed three important characteristics within the PGPB group: N 2 -fixation, siderophores, and indoles production.
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